HΔ24/FΔ30-pseudotyped LVs conserve the tropism of Edmonston MV strain. (A) SLAM and CD46 surface staining (open histograms) of the different cell lines (CHO, 293T, Jurkat, Raji, and B95a). Closed histograms represent staining with the isotype IgG control. (B) The different cell lines were transduced with a HΔ24/FΔ30-pseudotyped HIV-GFP vector at an MOI of 1 in the absence of antibody (no antibody) or after 2-hour incubation with an anti-CD46 antibody that blocks H binding to CD46 receptor or, as control, with anti-SLAM antibody prior to transduction. In the case of B95-a cells, a specific anti-SLAM antibody was used to block the SLAM receptor, whereas anti-CD46 antibody was used as a control prior to transduction. Cells were analyzed for GFP expression 72 hours after transduction by FACS analysis. Data shown here are representative of 4 independent experiments. (C) 293T cells were transduced with 2 different shRNA LVs to knock down hCD46 (293T-CD46^KD). Open histograms represent the expression of CD46 in 293T or 293T-CD46^KD. Closed histograms represent staining with the isotype IgG control. Both cell lines were subsequently transduced with HΔ24/FΔ30-pseudotyped HIV-GFP vector at MOI = 1. Dot blots represent the percentage of GFP⁺ cells 3 days after transduction, as analyzed by FACS.